34-METHYLENEDIOXYAMPHETAMINE Latest Revision August 16 2005

1 SYNONYMS

CFR 34-Methylenedioxyamphetamine

CAS Base 4764-17-4

Hydrochloride 6292-91-7

Other Names 34-Methylenedioxy-α-methyl-β-phenethylamine

34-Methylenedioxyphenylisopropylamine

alpha-Methyl-13-benzodioxole-5-ethanamine

alpha-Methyl-13-benzodioxole-5-ethanamine

Tenamfetamine

Adam

Love

EA-1299

MDA

2 CHEMICAL AND PHYSICAL DATA

21 CHEMICAL DATA

Form Chemical Formula Molecular Weight Melting Point (degC)

Base C10H13NO2 1792 Oil

Hydrochloride C10H13NO2middotHCl 2157 187-88

22 SOLUBILITY

Form A C E H M W

Base S S S S I

Hydrochloride SS S I S S

Sulfate I I I S

A = acetone C = chloroform E = ether H = hexane M = methanol and W = water VS = very soluble FS =

freely soluble S = soluble PS = sparingly soluble SS = slightly soluble VSS = very slightly soluble and I =

insoluble

3 SCREENING TECHNIQUES

31 COLOR TESTS

REAGENT COLOR PRODUCED

Marquis Purple to black

32 THIN LAYER CHROMATOGRAPHY

Visualization

Acidified potassium permanganate solution

COMPOUND

Relative Rf

System

TLC 5

System

TLC 6

34-methylenedioxyethylamphetamine 11 12

34-methylenedioxyamphetamine 10 10

34-methylenedioxydimethylamphetamine 09 12

34-methylenedioxymethamphetamine 08 06

33 GAS CHROMATOGRAPHY

Method MDA-GCS1

Instrument Gas Chromatograph operated in split mode with FID

Column 5 phenyl95 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperatures Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

Samples are to be dissolved in 41 chloroform methanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 061 acetaminophen 124

methamphetamine 067 caffeine 134

safrole 080 methyl stearate 149

C14 089 cocaine 161

MDA 10 (417 min) methyl eicosanoate 162

MDMA 105 tetraphenylethylene 178

MDEA 109 heroin 189

MMDA 112

Method MDA-GCS2

Instrument Gas Chromatograph operated in split mode with FID

Column 50 phenyl50 methyl silicone 30 m x 025 mm x 025 microm film thickness

Carrier gas Hydrogen at 20 mLmin

Temperatures Injector 260degC

Detector 270degC

Oven program

1) 90oC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

Samples are to be dissolved in 41 chloroform methanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 062 acetaminophen 131

methamphetamine 066 caffeine 138

safrole 080 methyl stearate 130

C14 069 cocaine 158

MDA 10 (484 min) methyl eicosanoate 141

MDMA 103 tetraphenylethylene 177

MDEA 105 heroin 203

MMDA 107

Method MDA-GCS3

Instrument Gas Chromatograph operated in split mode with FID

Column 5 phenyl95 methyl silicone 5 m x 025 mm x 025microm film

thickness

Carrier gas Hydrogen

1) Initial pressure of 25 psi for 010 min

2) Ramp to 50 psi at 341 psimin

3) Hold final pressure for 077 min

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature

2) Ramp to 100degC at 10degCmin

3) Hold final temperature for 060 min

Injection Parameters Split Ratio = 501 1 microL injected

EZFlash Parameters

1) Time 0 sec = 100degC

2) Time 6 sec = 110degC

3) Time 50 sec = 310degC

4) Time 96 sec = 310degC

Samples are to be dissolved in 41 chloroformmethanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 073 acetaminophen 119

methamphetamine 077 caffeine 127

safrole 085 methyl stearate 128

C14 092 cocaine 150

MDA 10 (479 min) methyl eicosanoate 140

MDMA 104 tetraphenylethylene 169

MDEA 107 heroin 188

MMDA 108

Method MDA-GCS4

Instrument Gas Chromatograph operated in split mode with FID

Column 1 phenyl99 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

COMPOUND RRT COMPOUND RRT

amphetamine 056 acetaminophen 127

methamphetamine 175 caffeine 136

safrole 077 methyl stearate 163

C14 097 cocaine 170

MDA 10 (41 min) methyl eicosanoate 178

MDMA 106 tetraphenylethylene 189

MDEA 112 heroin 201

MMDA 115

4 SEPARATION TECHNIQUES

5 QUANTITATIVE PROCEDURE

51 GAS CHROMATOGRAPHY

Method MDA-GCQ1

Internal Standard Stock Solution

10 mgmL n-butylamphetamine HCl in chloroform

Standard Solution Preparation

Prepare a standard solution of MDA HCl at 04 mgmL in chloroform

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

33 GAS CHROMATOGRAPHY

Method MDA-GCS1

Instrument Gas Chromatograph operated in split mode with FID

Column 5 phenyl95 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperatures Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

Samples are to be dissolved in 41 chloroform methanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 061 acetaminophen 124

methamphetamine 067 caffeine 134

safrole 080 methyl stearate 149

C14 089 cocaine 161

MDA 10 (417 min) methyl eicosanoate 162

MDMA 105 tetraphenylethylene 178

MDEA 109 heroin 189

MMDA 112

Method MDA-GCS2

Instrument Gas Chromatograph operated in split mode with FID

Column 50 phenyl50 methyl silicone 30 m x 025 mm x 025 microm film thickness

Carrier gas Hydrogen at 20 mLmin

Temperatures Injector 260degC

Detector 270degC

Oven program

1) 90oC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

Samples are to be dissolved in 41 chloroform methanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 062 acetaminophen 131

methamphetamine 066 caffeine 138

safrole 080 methyl stearate 130

C14 069 cocaine 158

MDA 10 (484 min) methyl eicosanoate 141

MDMA 103 tetraphenylethylene 177

MDEA 105 heroin 203

MMDA 107

Method MDA-GCS3

Instrument Gas Chromatograph operated in split mode with FID

Column 5 phenyl95 methyl silicone 5 m x 025 mm x 025microm film

thickness

Carrier gas Hydrogen

1) Initial pressure of 25 psi for 010 min

2) Ramp to 50 psi at 341 psimin

3) Hold final pressure for 077 min

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature

2) Ramp to 100degC at 10degCmin

3) Hold final temperature for 060 min

Injection Parameters Split Ratio = 501 1 microL injected

EZFlash Parameters

1) Time 0 sec = 100degC

2) Time 6 sec = 110degC

3) Time 50 sec = 310degC

4) Time 96 sec = 310degC

Samples are to be dissolved in 41 chloroformmethanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 073 acetaminophen 119

methamphetamine 077 caffeine 127

safrole 085 methyl stearate 128

C14 092 cocaine 150

MDA 10 (479 min) methyl eicosanoate 140

MDMA 104 tetraphenylethylene 169

MDEA 107 heroin 188

MMDA 108

Method MDA-GCS4

Instrument Gas Chromatograph operated in split mode with FID

Column 1 phenyl99 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

COMPOUND RRT COMPOUND RRT

amphetamine 056 acetaminophen 127

methamphetamine 175 caffeine 136

safrole 077 methyl stearate 163

C14 097 cocaine 170

MDA 10 (41 min) methyl eicosanoate 178

MDMA 106 tetraphenylethylene 189

MDEA 112 heroin 201

MMDA 115

4 SEPARATION TECHNIQUES

5 QUANTITATIVE PROCEDURE

51 GAS CHROMATOGRAPHY

Method MDA-GCQ1

Internal Standard Stock Solution

10 mgmL n-butylamphetamine HCl in chloroform

Standard Solution Preparation

Prepare a standard solution of MDA HCl at 04 mgmL in chloroform

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Method MDA-GCS2

Instrument Gas Chromatograph operated in split mode with FID

Column 50 phenyl50 methyl silicone 30 m x 025 mm x 025 microm film thickness

Carrier gas Hydrogen at 20 mLmin

Temperatures Injector 260degC

Detector 270degC

Oven program

1) 90oC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

Samples are to be dissolved in 41 chloroform methanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 062 acetaminophen 131

methamphetamine 066 caffeine 138

safrole 080 methyl stearate 130

C14 069 cocaine 158

MDA 10 (484 min) methyl eicosanoate 141

MDMA 103 tetraphenylethylene 177

MDEA 105 heroin 203

MMDA 107

Method MDA-GCS3

Instrument Gas Chromatograph operated in split mode with FID

Column 5 phenyl95 methyl silicone 5 m x 025 mm x 025microm film

thickness

Carrier gas Hydrogen

1) Initial pressure of 25 psi for 010 min

2) Ramp to 50 psi at 341 psimin

3) Hold final pressure for 077 min

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature

2) Ramp to 100degC at 10degCmin

3) Hold final temperature for 060 min

Injection Parameters Split Ratio = 501 1 microL injected

EZFlash Parameters

1) Time 0 sec = 100degC

2) Time 6 sec = 110degC

3) Time 50 sec = 310degC

4) Time 96 sec = 310degC

Samples are to be dissolved in 41 chloroformmethanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 073 acetaminophen 119

methamphetamine 077 caffeine 127

safrole 085 methyl stearate 128

C14 092 cocaine 150

MDA 10 (479 min) methyl eicosanoate 140

MDMA 104 tetraphenylethylene 169

MDEA 107 heroin 188

MMDA 108

Method MDA-GCS4

Instrument Gas Chromatograph operated in split mode with FID

Column 1 phenyl99 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

COMPOUND RRT COMPOUND RRT

amphetamine 056 acetaminophen 127

methamphetamine 175 caffeine 136

safrole 077 methyl stearate 163

C14 097 cocaine 170

MDA 10 (41 min) methyl eicosanoate 178

MDMA 106 tetraphenylethylene 189

MDEA 112 heroin 201

MMDA 115

4 SEPARATION TECHNIQUES

5 QUANTITATIVE PROCEDURE

51 GAS CHROMATOGRAPHY

Method MDA-GCQ1

Internal Standard Stock Solution

10 mgmL n-butylamphetamine HCl in chloroform

Standard Solution Preparation

Prepare a standard solution of MDA HCl at 04 mgmL in chloroform

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature

2) Ramp to 100degC at 10degCmin

3) Hold final temperature for 060 min

Injection Parameters Split Ratio = 501 1 microL injected

EZFlash Parameters

1) Time 0 sec = 100degC

2) Time 6 sec = 110degC

3) Time 50 sec = 310degC

4) Time 96 sec = 310degC

Samples are to be dissolved in 41 chloroformmethanol and filtered

COMPOUND RRT COMPOUND RRT

amphetamine 073 acetaminophen 119

methamphetamine 077 caffeine 127

safrole 085 methyl stearate 128

C14 092 cocaine 150

MDA 10 (479 min) methyl eicosanoate 140

MDMA 104 tetraphenylethylene 169

MDEA 107 heroin 188

MMDA 108

Method MDA-GCS4

Instrument Gas Chromatograph operated in split mode with FID

Column 1 phenyl99 methyl silicone 30 m x 025 mm x 025 microm film

thickness

Carrier gas Hydrogen at 20 mLmin

Temperature Injector 260degC

Detector 270degC

Oven program

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

COMPOUND RRT COMPOUND RRT

amphetamine 056 acetaminophen 127

methamphetamine 175 caffeine 136

safrole 077 methyl stearate 163

C14 097 cocaine 170

MDA 10 (41 min) methyl eicosanoate 178

MDMA 106 tetraphenylethylene 189

MDEA 112 heroin 201

MMDA 115

4 SEPARATION TECHNIQUES

5 QUANTITATIVE PROCEDURE

51 GAS CHROMATOGRAPHY

Method MDA-GCQ1

Internal Standard Stock Solution

10 mgmL n-butylamphetamine HCl in chloroform

Standard Solution Preparation

Prepare a standard solution of MDA HCl at 04 mgmL in chloroform

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

1) 90degC initial temperature for 10 min

2) Ramp to 295degC at 30degmin

3) Hold final temperature for 26 min

Injection Parameters Split Ratio = 501 1 microL injected

COMPOUND RRT COMPOUND RRT

amphetamine 056 acetaminophen 127

methamphetamine 175 caffeine 136

safrole 077 methyl stearate 163

C14 097 cocaine 170

MDA 10 (41 min) methyl eicosanoate 178

MDMA 106 tetraphenylethylene 189

MDEA 112 heroin 201

MMDA 115

4 SEPARATION TECHNIQUES

5 QUANTITATIVE PROCEDURE

51 GAS CHROMATOGRAPHY

Method MDA-GCQ1

Internal Standard Stock Solution

10 mgmL n-butylamphetamine HCl in chloroform

Standard Solution Preparation

Prepare a standard solution of MDA HCl at 04 mgmL in chloroform

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Sample Preparation

Accurately weigh out 50 mg of sample into a 25 mL volumetric flask and dilute with water to the mark Take a

2 mL aliquot of the solution add 3 mL of IS and 2-3 mL of 05 M KOH Shake Separate layers and extract

twice more with 2-3 mL each time of chloroform Combine fractions ndash dry over sodium sulfate and dilute to

total volume of 10 mL Treat the standard in the same fashion as the sample prior to analysis

Instrument Gas Chromatograph operated in split mode with FID

Column DB-1 30 m x 025 mm x 025 m film thickness

Carrier gas Hydrogen at 13 mLmin

Make-Up gas Nitrogen at 400 mLmin

Temperatures Injector 230degC

Detector 280degC

Oven Program

1) 150degC initial temperature 2 min

2) Ramp to 190degC at 5degCmin

3) Hold final temperature for 10 min

Injection Parameters Split Ratio = 201 1microL injected

Typical Retention Time 34-MDA 213 min

N-Butylamphetamine HCl 189 min

Linear Range 006 mgmL to 20 mgmL

Repeatability RSD less than 20

Correlation Coefficient 09998

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT COMPOUND RRT

methylbenzylamine 040 salicylamide 089 caffeine 214

P2P 044 methyl paraben 089 antipyrine 230

amphetamine 045 penicillin 090 benzphetamine 235

nicotinic acid 048 phenmetrazine 093 diphenhydramine 246

phentermine 048 MDP-2-P 097 aminopyrine 260

methamphetamine 051 phendimetrazine 099 doxylamine 266

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

COMPOUND RRT COMPOUND RRT COMPOUND RRT

fenfluramine 055 MDA 10 (213 min) phthalic acid 282

ethylamphetamine 056 MDMA 115 palmitic acid 282

dimethylamphetamine 058 aminorex 121 dipyrone 289

safrole 063 methyl aminorex 121 procaine 306

salicylic acid 064 MDEA 130 eicosane 307

cathine 068 ibuprofen 143 dextromethorphan 359

phenylpropanolamine 068 MBDB 145 strychnine 365

methcathinone 069 hexadecane 147 amitriptyline 384

nicotinamide 071 guaifenesin 150 scopolamine 427

chlorpheniramine 075 acetaminophen 155 tetracosane 458

chloroephedrine 076 MMDA 159 chlordiazepoxide 496

pseudoephedrine 076 phenacetin 162 quinine 602

ephedrine 078 chloromdma 171

butylamphetamine 089 methylphenidate 187

52 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Method MDA-LCQ1

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 10 mgmL using water Filter

with a 045-microm filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with water If necessary dilute the

sample so the final concentration approximates the standard concentration or falls within the linear range Filter

sample with a 045-microm filter

Instrument High performance liquid chromatograph equipped with diode array

Column 5 microm ODS 40 mm x 125 mm

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Flow 10 mLmin

Injection Volume 1 microL

Buffer 4000 mL distilled water 30 mL phosphoric acid 10 g sodium

hydroxide and 80 mL hexylamine at pH 25

Mobile Phase Buffer acetonitrile 8515 with gradient to 6040 over 6 min

Typical Retention Time 34-MDA 163 min

Linear Range 01338 - 428 mgmL

Repeatability RSD less than 02

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT

Nicotinamide 062

MDA 100

MDMA 116

MDEA 141

Method MDA-LCQ2

Standard Solution Preparation

Prepare a standard solution of MDA HCl at approximately 05 mgmL using water buffer or methanol

Sample Preparation

Accurately weigh an amount of sample into an appropriate volumetric or Erlenmeyer flask and dilute so that the

final MDA HCl concentration is approximately that of the standard solution

Instrument High performance liquid chromatograph equipped with diode array

Column 5 Phenomenex Luna 150 mm x 32 mm at 35 C

Detector UV 210 nm

Flow 10 mLmin

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Injection Volume 5 microL

Buffer 4000 mL distilled water 225 mL phosphoric acid adjust to pH 22-23

with triethanolamine (approx 22 mL)

Mobile Phase Bufferacetonitrile 9010

Typical Retention Time 34-MDA 98 min

Linear Range 005 mgmL - 2 mgmL

Repeatability RSD less than 05

Correlation Coefficient 09999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

phenylpropanolamine 065 methyl aminorex 109

methyl benzylamine 067 phentermine 110

cathine 068 chloroephedrine 111

doxylamine 074 ethylamphetamine 113

dipyrone 075 MDP-2-P 113

ephedrine 077 lidocaine 115

methcathinone 079 caffeine 116

pseudoephedrine 079 MDEA 122

amphetamine 090 phenyl-2-naphthalene 122

theophylline 094 phthalic acid 122

methapyrilene 096 strychnine 123

phenmetrazine 096 MMDA 126

methamphetamine 098 MBDB 134

phendimetrazine 098 salicylamide 137

scopolamine 099 acetaminophen 139

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

COMPOUND RRT COMPOUND RRT

MDA 100 (98 min) chloro-mdma 147

quinine 103 methylphenidate 148

saccharin 103 glycerol glycolate 159

dimethylamphetamine 104 chlorpheniramine 167

MDMA 108 aspirin 195

chlordiazepoxide 201

53 CAPILLARY ELECTROPHORESIS

Method MDA-CEQ1

Internal Standard Stock Solution

050 mgmL n-butylamphetamine HCl in 50 mM sodium phosphate at pH 70

Standard Solution Preparation

Accurately weigh and prepare a standard solution of MDA HCl at approximately 08 mgmL using the internal

standard stock solution Filter with 045 micron filter

Sample Preparation

Accurately weigh an amount of sample into a volumetric flask and dilute with internal standard stock solution

If necessary dilute the sample so the final concentration approximates the standard concentration or falls within

the linear range Filter sample with 045-micron filter

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Mode Cyclodextrin system analysis

Column 49 cm x 52 microm fused silica capillary

Run Buffer 10 mM gamma-highly sulfated cyclodextrin in 50 mM sodium

phosphate at pH 70

Detector UV 210 nm

Voltage 12 kV

Temperature 30degC air cooled

Injection 1 s hydrodynamic at 50 mbars

Run Time 17 min

Rinse Time 20 min

Linear Range 002 - 100 mgmL

Repeatability RSD of area less than 15

Correlation Coefficient 0999

Accuracy Error less than 5

COMPOUND RRT COMPOUND RRT

d-methamphetamine 083 l-pseudoephedrine 123

d-ephedrine 093 MDMA 125

MDA 10 (883 min) l-ephedrine 136

cathine 106 MDEA 140

l-methamphetamine 111 MDMA 148

d-amphetamine 118 l-norephedrine 165

d-norephedrine 118 l-amphetamine 168

d-pseudoephedrine 121 MDA 170

MDEA 186

Method MDA-CEQ2

Internal Standard Stock Solution

10 mgmL n-Butylamphetamine HCl in 38 mM phosphate buffer (pH 25)

Standard Solution Preparation

Weigh an appropriate amount of standard MDA HCl into a volumetric flask to obtain a final concentration of

approximately 008 mgmL Pipette an appropriate amount of internal standard solution to obtain a final

concentration of 01 mgmL Dilute to volume with injection solvent Filter approximately 10 mL of solution

with regenerated cellulose 045 microm 25mm filter into a 20 mL glass vial (Agilent part number 5182-0567)

Make sure there are no air bubbles on bottom of glass vial Cap vial with polypropylene cap (Agilent part

number 5182-9697)

Sample Preparation

Weigh an appropriate amount of sample into a volumetric flask so that the final phenethylamine concentration

is approximately that of the standard solution Pipette appropriate amount of internal standard solution to obtain

a final concentration of 01 mgmL Dilute to volume with injection solvent Sonicate for 15 minutes Filter

approximately 10 mL of solution with regenerated cellulose 045 m 25mm filter into a 20 mL glass vial

(Agilent part number 5182-0567) Make sure there are no air bubbles on bottom of glass vial Cap vial with

polypropylene cap (Agilent part number 5182-9697)

Mode HP 3D instrument operated in CE mode

Cyclodextrin system analysis

Column 50 microm id x 322 cm

(237 cm length to detector)

Run Buffer Celixir Reagent B pH 25 (MicroSolv CE)

Detector Diode-array

Voltage 10 kV

Temperature 15degC

Injection 50 mbar x 2 s followed by water at 35 mbar

Run Time 6 min

Rinse Time 2 min

Linear Range 002 ndash 100 mgmL

Repeatability RSD lt16

Correlation Coefficient 10

Accuracy Error less than 32

COMPOUND RRT COMPOUND RRT

doxylamine 0765 ephedrine 0932

chlorpheniramine 0784 phenylephrine 0951

quinine 0804 MDEA 0961

-phenethylamine 0807 ketamine 0962

chlorquinine 0812 phenyltoxolamine 0971

nicotinamide 0836 n-butylamphetamine 10 (46 min)

amphetamine 0868 dextromethorphan 100

methamphetamine 0883 lidocaine 103

procaine 0883 benzocaine 125

MDA 0900 acetaminophen 211

norpseudoephedrine 0906 caffeine 214

MDMA 0914 guaifenesin 214

norephedrine 0917 P2P 224

pseudoephedrine 0919 DMSO (neutral marker) 240

tetracaine 0927 aspirin 271

54 NUCLEAR MAGNETIC RESONANCE

Method NMR-1-1

Solvents Deuterium Oxide (D2O) containing DSS or TSP for 0 PPM reference

Internal Standard Stock Solution (ISSS) 5 mgmL Maleic Acid (accurately weighed) in deuterium oxide (D2O)

containing DSS or TSP for 0 ppm reference

Sample Preparation Accurately weigh an amount of sample usually 10-30 mg into a capped test tube and

accurately add a volume normally 10 mL of the ISSS Vortex the sample for several seconds If insolubles

are present add 10 mL D2O (not containing maleic acid or the reference compound) vortex and sonicate 15

minutes Vortex the sample and filter if necessary Place in NMR sample tube

Instrument Varian Mercury 400 MHz with proton detection probe NMR Spectrometer

Pulse Width 10 micros or 90deg (whichever is less)

Spectral Width (SW) at least containing ndash3 ppm through 13 ppm

Number of Scans multiple of 4 (greater to enhance SN)

Delay between Pulses 45 s

Shimming automatic gradient shimming of Z1-4 shims

Total run timesample 6 min (NT = 4) -14 min (NT = 16)

Uniformity within spectral width 03 RSD (-06 to 114 PPM)

Run Time 6 min

Linear Range 06 mgmL - 67 mgmL

Repeatability lt4

Correlation Coefficient 10

Accuracy lt3

6 QUALITATIVE DATA

See spectra on the following pages for Mass Spectrometry FT-IR Vapor Phase IR FT-Raman and Nuclear

Magnetic Resonance

7 REFERENCES

Coates J and Reffner J Visualization of Micro-ATR Infrared Spectroscopy Spectroscopy Vol 14 4

April 1999

Clarke EGC Isolation and Identification of Drugs 2nd Edition The Pharmaceutical Press 1986

Galichat Laurent Y Clarkersquos Analysis of Drugs and Poisons Volume 2 p 1256 Pharmaceutical Press 2004

Budavari S The Merck Index 13th Edition Merck and Co Inc 2001

8 ADDITIONAL RESOURCES

Forendex

Wikipedia

Optical isomers standards for these compounds were unavailable

No Data Available

Correlation Coefficient 10

Accuracy Error less than 32

COMPOUND RRT COMPOUND RRT

doxylamine 0765 ephedrine 0932

chlorpheniramine 0784 phenylephrine 0951

quinine 0804 MDEA 0961

-phenethylamine 0807 ketamine 0962

chlorquinine 0812 phenyltoxolamine 0971

nicotinamide 0836 n-butylamphetamine 10 (46 min)

amphetamine 0868 dextromethorphan 100

methamphetamine 0883 lidocaine 103

procaine 0883 benzocaine 125

MDA 0900 acetaminophen 211

norpseudoephedrine 0906 caffeine 214

MDMA 0914 guaifenesin 214

norephedrine 0917 P2P 224

pseudoephedrine 0919 DMSO (neutral marker) 240

tetracaine 0927 aspirin 271

54 NUCLEAR MAGNETIC RESONANCE

Method NMR-1-1

Solvents Deuterium Oxide (D2O) containing DSS or TSP for 0 PPM reference

Internal Standard Stock Solution (ISSS) 5 mgmL Maleic Acid (accurately weighed) in deuterium oxide (D2O)

containing DSS or TSP for 0 ppm reference

Sample Preparation Accurately weigh an amount of sample usually 10-30 mg into a capped test tube and

accurately add a volume normally 10 mL of the ISSS Vortex the sample for several seconds If insolubles

are present add 10 mL D2O (not containing maleic acid or the reference compound) vortex and sonicate 15

minutes Vortex the sample and filter if necessary Place in NMR sample tube

Instrument Varian Mercury 400 MHz with proton detection probe NMR Spectrometer

Pulse Width 10 micros or 90deg (whichever is less)

Spectral Width (SW) at least containing ndash3 ppm through 13 ppm

Number of Scans multiple of 4 (greater to enhance SN)

Delay between Pulses 45 s

Shimming automatic gradient shimming of Z1-4 shims

Total run timesample 6 min (NT = 4) -14 min (NT = 16)

Uniformity within spectral width 03 RSD (-06 to 114 PPM)

Run Time 6 min

Linear Range 06 mgmL - 67 mgmL

Repeatability lt4

Correlation Coefficient 10

Accuracy lt3

6 QUALITATIVE DATA

See spectra on the following pages for Mass Spectrometry FT-IR Vapor Phase IR FT-Raman and Nuclear

Magnetic Resonance

7 REFERENCES

Coates J and Reffner J Visualization of Micro-ATR Infrared Spectroscopy Spectroscopy Vol 14 4

April 1999

Clarke EGC Isolation and Identification of Drugs 2nd Edition The Pharmaceutical Press 1986

Galichat Laurent Y Clarkersquos Analysis of Drugs and Poisons Volume 2 p 1256 Pharmaceutical Press 2004

Budavari S The Merck Index 13th Edition Merck and Co Inc 2001

8 ADDITIONAL RESOURCES

Forendex

Wikipedia

Optical isomers standards for these compounds were unavailable

No Data Available

Internal Standard Stock Solution (ISSS) 5 mgmL Maleic Acid (accurately weighed) in deuterium oxide (D2O)

containing DSS or TSP for 0 ppm reference

Sample Preparation Accurately weigh an amount of sample usually 10-30 mg into a capped test tube and

accurately add a volume normally 10 mL of the ISSS Vortex the sample for several seconds If insolubles

are present add 10 mL D2O (not containing maleic acid or the reference compound) vortex and sonicate 15

minutes Vortex the sample and filter if necessary Place in NMR sample tube

Instrument Varian Mercury 400 MHz with proton detection probe NMR Spectrometer

Pulse Width 10 micros or 90deg (whichever is less)

Spectral Width (SW) at least containing ndash3 ppm through 13 ppm

Number of Scans multiple of 4 (greater to enhance SN)

Delay between Pulses 45 s

Shimming automatic gradient shimming of Z1-4 shims

Total run timesample 6 min (NT = 4) -14 min (NT = 16)

Uniformity within spectral width 03 RSD (-06 to 114 PPM)

Run Time 6 min

Linear Range 06 mgmL - 67 mgmL

Repeatability lt4

Correlation Coefficient 10

Accuracy lt3

6 QUALITATIVE DATA

See spectra on the following pages for Mass Spectrometry FT-IR Vapor Phase IR FT-Raman and Nuclear

Magnetic Resonance

7 REFERENCES

Coates J and Reffner J Visualization of Micro-ATR Infrared Spectroscopy Spectroscopy Vol 14 4

April 1999

Clarke EGC Isolation and Identification of Drugs 2nd Edition The Pharmaceutical Press 1986

Galichat Laurent Y Clarkersquos Analysis of Drugs and Poisons Volume 2 p 1256 Pharmaceutical Press 2004

Budavari S The Merck Index 13th Edition Merck and Co Inc 2001

8 ADDITIONAL RESOURCES

Forendex

Wikipedia

Optical isomers standards for these compounds were unavailable

No Data Available

Galichat Laurent Y Clarkersquos Analysis of Drugs and Poisons Volume 2 p 1256 Pharmaceutical Press 2004

Budavari S The Merck Index 13th Edition Merck and Co Inc 2001

8 ADDITIONAL RESOURCES

Forendex

Wikipedia

Optical isomers standards for these compounds were unavailable

No Data Available

Mass Spectrum (EI) 34-Methylenedioxyamphetamine HCl Lot A150B

FTIR (One bounce - ATR) 34-Methylenedioxyamphetmaine HCl Lot A150B

4 cm-1

resolution 32 scans

17

80863103337

123768

148695

290079

70

72

74

76

78

80

82

84

86

88

90

92

94

96

T

500 1000 1500 2000 2500 3000 3500 4000

Wavenumbers (cm-1)

FTIR (One bounce ndash ATR) 34-Methylenedioxyamphetamine sulfate Lot X-19LDS

4 cm-1

resolution 32 scans

GCIRD 34-Methylenedioxyamphetamine Lot A150B

4 cm-1

resolution 2 mgmL in chloroform

45312

62064

102923

108235

125805

148277

68

70

72

74

76

78

80

82

84

86

88

90

92

94

96

T

500 1000 1500 2000 2500 3000 3500

Wavenumbers (cm-1)

FT-RAMAN 34-Methylenedioxyamphetamine HCl Lot A150B

4 cm-1

resolution 32 scans

FT-RAMAN 34-Methylenedioxyamphetamine sulfate Lot X-19-LDS

4 cm-1

resolution 32 scans

DEA

52

93

81

13

12

56

8

13

75

3

29

32

4

30

83

6

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

DEA

71

32

81

13

97

48

29

28

3

30

67

3

02

04

06

08

10

12

14

16

18

20

22

24

26

28

30

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

1H NMR 34-Methylenedioxyamphetamine HCl Lot A150B

deuterium oxide maleic acid as internal standard 400 MHz

ppm 688 (d J=79 Hz 1 H) 683 (d J=14 Hz 1 H) 679 (dd J=79 14

Hz 1 H) 596 (s 2 H) 359 (tq J=72 67 Hz 1 H) 287 (d J=72 Hz 2

H) 132 (d J=67 Hz 3 H)

13

C NMR 34 Methylenedioxyamphetamine HCl Lot A 150B

deuterium oxide maleic acid as internal standard

FTIR (One bounce ndash ATR) 34-Methylenedioxyamphetamine sulfate Lot X-19LDS

4 cm-1

resolution 32 scans

GCIRD 34-Methylenedioxyamphetamine Lot A150B

4 cm-1

resolution 2 mgmL in chloroform

45312

62064

102923

108235

125805

148277

68

70

72

74

76

78

80

82

84

86

88

90

92

94

96

T

500 1000 1500 2000 2500 3000 3500

Wavenumbers (cm-1)

FT-RAMAN 34-Methylenedioxyamphetamine HCl Lot A150B

4 cm-1

resolution 32 scans

FT-RAMAN 34-Methylenedioxyamphetamine sulfate Lot X-19-LDS

4 cm-1

resolution 32 scans

DEA

52

93

81

13

12

56

8

13

75

3

29

32

4

30

83

6

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

DEA

71

32

81

13

97

48

29

28

3

30

67

3

02

04

06

08

10

12

14

16

18

20

22

24

26

28

30

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

1H NMR 34-Methylenedioxyamphetamine HCl Lot A150B

deuterium oxide maleic acid as internal standard 400 MHz

ppm 688 (d J=79 Hz 1 H) 683 (d J=14 Hz 1 H) 679 (dd J=79 14

Hz 1 H) 596 (s 2 H) 359 (tq J=72 67 Hz 1 H) 287 (d J=72 Hz 2

H) 132 (d J=67 Hz 3 H)

13

C NMR 34 Methylenedioxyamphetamine HCl Lot A 150B

deuterium oxide maleic acid as internal standard

FT-RAMAN 34-Methylenedioxyamphetamine HCl Lot A150B

4 cm-1

resolution 32 scans

FT-RAMAN 34-Methylenedioxyamphetamine sulfate Lot X-19-LDS

4 cm-1

resolution 32 scans

DEA

52

93

81

13

12

56

8

13

75

3

29

32

4

30

83

6

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

DEA

71

32

81

13

97

48

29

28

3

30

67

3

02

04

06

08

10

12

14

16

18

20

22

24

26

28

30

Int

500 1000 1500 2000 2500 3000 3500

Raman shift (cm-1)

1H NMR 34-Methylenedioxyamphetamine HCl Lot A150B

deuterium oxide maleic acid as internal standard 400 MHz

ppm 688 (d J=79 Hz 1 H) 683 (d J=14 Hz 1 H) 679 (dd J=79 14

Hz 1 H) 596 (s 2 H) 359 (tq J=72 67 Hz 1 H) 287 (d J=72 Hz 2

H) 132 (d J=67 Hz 3 H)

13

C NMR 34 Methylenedioxyamphetamine HCl Lot A 150B

deuterium oxide maleic acid as internal standard

1H NMR 34-Methylenedioxyamphetamine HCl Lot A150B

deuterium oxide maleic acid as internal standard 400 MHz

ppm 688 (d J=79 Hz 1 H) 683 (d J=14 Hz 1 H) 679 (dd J=79 14

Hz 1 H) 596 (s 2 H) 359 (tq J=72 67 Hz 1 H) 287 (d J=72 Hz 2

H) 132 (d J=67 Hz 3 H)

13

C NMR 34 Methylenedioxyamphetamine HCl Lot A 150B

deuterium oxide maleic acid as internal standard